Assessment of agonist- and cell-mediated responses in airway microsections by computerized videomicrometry.
Assessment of agonist- and cell-mediated responses in airway microsections by computerized videomicrometry.
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通过计算机视频显微测量法评估气道显微切片中激动剂和细胞介导的反应。
DOI：
10.1152/ajplung.1995.268.3.l519
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Leff,AR
中科院分区：
文献类型：
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作者：
Galens,S;Munoz,NM;Rabe,KF;Herrnreiter,A;Mayer,D;Morton,B;McAllister,K;Leff,AR
本研究的目的是开发一种方法，用于实时测量药理学和生理学干预期间气道微外植体中管腔面积的变化。在杀死豚鼠后，切下气管环（1至2 mm厚）并置于300微升的腔室中。使用计算机化视频显微测量法计算气道腔的面积作为像素数。在29个上皮完整的气道中，10（-3）M乙酰胆碱（ACh）导致管腔面积减少38.1 ± 2.80%（P < 0.001 vs. 10（-9）M）。在4只豚鼠的34份制备物中也证实了自发张力;在单独缓冲液中孵育60分钟后，面积减少17.0 +/- 1.45%，可被10（-5）M吲哚美辛完全阻断（P = 0.01）。10（-6）M沙丁胺醇可完全逆转≤ 10（-6）M ACh引起的管腔狭窄（P = 0.002）。加入100，000个活化的人嗜酸性粒细胞导致管腔面积减少24.7 +/- 4.41%，而非活化细胞减少7.24 +/- 5.51%（P = 0.048）。我们展示了一种实时的方法，用于评估的营养不良的变化，在气道口径，利用显微镜下的呼吸道，并允许使用极少量的分离的细胞，以实现生理激活。浓度-反应特征和自发性音调与大腔室制剂相似，并且狭窄可通过β 2-肾上腺素受体激活逆转。
The objective of this investigation was to develop a method for real-time measurement of changes in luminal area in microexplants of airways during pharmacological and physiological interventions. After guinea pigs were killed, tracheal rings (1- to 2-mm thick) were excised and placed in 300-microliters chambers. The area of the airway lumen was calculated as pixel number with the use of computerized videomicrometry. In 29 epithelium-intact airways, 10(-3) M acetylcholine (ACh) caused decrease in luminal area of 38.1 +/- 2.80% (P < 0.001 vs. 10(-9) M). Spontaneous tone also was demonstrated in 34 preparations from 4 guinea pigs; decrease in area of 17.0 +/- 1.45% after 60-min incubation in buffer alone was blocked completely by 10(-5) M indomethacin (P = 0.01). Luminal narrowing caused by < or = 10(-6) M ACh was reversed completely by 10(-6) M albuterol (P = 0.002). Addition of 100,000 activated human eosinophils caused 24.7 +/- 4.41% decrease in luminal area vs. 7.24 +/- 5.51% for nonactivated cells (P = 0.048). We demonstrate a real-time method for the assessment of auxotonic changes in airway caliber that utilizes microsections of explanted airways and permits the use of extremely small numbers of isolated cells to achieve physiological activation. Concentration-response characteristics and spontaneous tone are similar to those of large chamber preparations, and narrowing is reversed by beta 2-adrenoceptor activation.